A breeding method of chou's parasitoid wasps based on female and male separation inoculation of pupae of antheraea pernyi

Abstract

The application discloses a kind of based on tussah pupae sex separation inoculation Zhou's fig wasp breeding method, first with shell tussah pupae airing after stripping cocoon, artificial sex separation and synchronous elimination damaged pupae and disease body pupae, again by washing disinfection, healthy female, male tussah pupae are respectively loaded into flat shallow container.Inoculation according to the order of female first and then male, with the average amount of single head of mixed inoculation as reference, female single head is higher than the average value, and male single head is lower than the average value.Inoculation container is closed and compacted, and is moved into the breeding room without light and micro-flow, and parasitic culture is carried out by adjusting temperature and humidity.Subsequently, the parasitic pupae are placed in the development interval for further development, and then transferred to low-temperature cold storage after gradient cooling for standby.The application significantly improves the parasitization rate of tussah pupae, reduces the rate of rotten pupae, saves the bee resource, shortens the adult emergence period, and realizes the efficient breeding of Zhou's fig wasp.

Description

Technical Field

[0001] This invention relates to the field of biological control and insect breeding technology, specifically to a method for breeding *Tetranychus zhouensis* based on the separation and inoculation of male and female *tussah silkworm* pupae. This method significantly improves the parasitism rate and bee utilization rate, reduces the rate of rotten pupae, and achieves efficient and large-scale breeding of *Tetranychus zhouensis* by separating male and female *tussah silkworm* pupae and adjusting the amount of bee colonies released according to the differences in the biological characteristics of male and female pupae. Background Technology

[0002] *Chouioia cunea* is an important parasitic natural enemy insect, widely used in the biological control of forestry and agricultural pests. Due to its large size and rich nutritional content, the tussah silkworm pupa (*Antherae aperrinyi*) is widely used as a substitute host for breeding *Chouioia cunea*. Traditional breeding methods for *Chouioia cunea* typically involve inoculating mixed-sex tussah silkworm pupae, placing both female and male pupae in the same breeding container, and then introducing wasp larvae at a specific ratio.

[0003] However, traditional methods have the following significant drawbacks: First, due to the natural differences in body size, epidermal thickness, and nutrient composition between female and male tussah silkworm pupae, their attractiveness and carrying capacity to *Paragonimus westermani* species differ. During mixed inoculation, the bee species tend to parasitize pupae of one sex, resulting in a low parasitism rate for the other sex and making it difficult to improve the overall parasitism rate. Second, mixed inoculation cannot optimize the bee species release quantity based on the characteristics of pupae of different sexes, easily leading to bee species waste or insufficient parasitism, which in turn increases the rate of rotten pupae, affecting the quality and quantity of emerging bees. Furthermore, traditional methods lack effective removal of diseased and damaged pupae and standardized disinfection procedures, further exacerbating rotten pupae, shortening the adult emergence cycle, and reducing overall breeding efficiency. Therefore, the industry urgently needs a new *Paragonimus westermani* breeding technology that can overcome the above-mentioned drawbacks and improve breeding efficiency and quality. Summary of the Invention

[0004] To address the shortcomings of existing technologies, this invention provides a method for breeding *Parasiticus zhoui* based on the separate inoculation of male and female *tussah silkworm* pupae. The core of this method lies in separating the male and female *tussah silkworm* pupae before inoculation and placing them separately into breeding containers. Then, based on the different characteristics of female and male *tussah silkworm* pupae, a larger quantity of *Parasiticus zhoui* is placed in the container for female pupae, and a smaller quantity is placed in the container for male pupae. This method can significantly improve the parasitism rate of *tussah silkworm* pupae, reduce the rate of pupal rot, conserve the number of *Parasiticus zhoui*, and shorten the adult emergence period.

[0005] Specifically, the technical solution of the present invention is as follows: A method for breeding *Pterocarya zhouensis* based on the separate inoculation of male and female *Tetraphyta tussah* pupae includes the following steps: S1. Separate the male and female tussah silkworm pupae to obtain female and male tussah silkworm pupae; S2. Place the female and male tussah silkworm pupae into separate breeding containers; S3. A first number of *Apis zhouensis* species are placed into the breeding container containing the female *tussah silkworm* pupae, and a second number of *Apis zhouensis* species are placed into the breeding container containing the male *tussah silkworm* pupae; wherein, the number of species placed for a single female *tussah silkworm* pupae is greater than the number of species placed for a single male *tussah silkworm* pupae. S4. Seal the inoculated breeding container and transfer it to the breeding room for parasitic culture.

[0006] Furthermore, prior to step S1, a pretreatment of the silkworm pupae with their shells is included: the silkworm pupae with their shells are placed indoors to dry for 2-3 hours until the cocoon shells are dry and hardened before peeling them off to obtain the naked silkworm pupae. This pretreatment step dehydrates and hardens the cocoon shells, making them easier to peel manually, while reducing mechanical damage to the pupae during the peeling process, thus providing intact and healthy pupae for subsequent male-female separation.

[0007] Furthermore, during the process of separating male and female tussah silkworm pupae, damaged and diseased pupae are simultaneously removed, retaining only healthy and intact tussah silkworm pupae. This screening step can promptly remove individuals that may carry pathogens or are already damaged, preventing the spread of disease in the breeding population and significantly reducing the subsequent rate of rotten pupae.

[0008] Furthermore, before placing the female and male tussah silkworm pupae into separate breeding containers, a cleaning and disinfection step is included: the separated female and male tussah silkworm pupae are washed with clean water, dried, and then surface-disinfected with a specific disinfectant (e.g., 0.1% sodium hypochlorite solution). This step effectively removes dirt and microorganisms from the surface of the pupae, further reducing the risk of disease infection during parasitism.

[0009] Further, step S3, which involves placing a first number of *Ariocarpus zhouensis* larvae into the breeding container containing the female *Tetracentron sinense* pupae and a second number of *Ariocarpus zhouensis* larvae into the breeding container containing the male *Tetracentron sinense* pupae, specifically includes: Following the inoculation order of females first and then males, the Zhou's biting wasp species were placed into the corresponding breeding containers. Taking the average number of wasp species per tussah silkworm pupa during mixed inoculation as a reference, the number of wasp species per female tussah silkworm pupa was higher than the average number, while the number of wasp species per male tussah silkworm pupa was lower than the average number.

[0010] The scientific basis for this differentiated deployment strategy is as follows: female tussah silkworm pupae are typically larger and richer in nutrients, enabling them to support more bee species in parasitism and development. Therefore, appropriately increasing the number of bee species deployed can improve the utilization rate and parasitism efficiency of female pupae. Male tussah silkworm pupae, on the other hand, are relatively smaller, and excessive parasitism can lead to intensified resource competition, poor individual development, and even pupa rot. Therefore, appropriately reducing the number of bee species deployed ensures the healthy development of parasitic bees while conserving bee resources. The female-first, male-later inoculation order helps bee species prioritize the use of more suitable female hosts, further optimizing the parasitism effect.

[0011] Furthermore, the parasitic culture step specifically includes: sealing and compacting the inoculated breeding container, transferring it to a breeding room with no light and slight air circulation, controlling the temperature in the breeding room to be 25℃-28℃ and the relative humidity to be 60%-70%, and storing it for 36-48 hours to allow the Zhou's parasitic wasp species to complete the parasitism of the tussah silkworm pupae. These temperature and humidity conditions and storage time have been optimized to provide the best environment for the initial parasitism of Zhou's parasitic wasps. The sealing and compacting operation prevents the wasp species from escaping, while the no-light and slight-circulation conditions simulate a natural parasitic environment, which is conducive to the successful egg-laying of the wasps.

[0012] Furthermore, after the parasitic culture is completed, a developmental step is included: the parasitized silkworm pupae are placed in a single layer and moved to a developmental chamber, where they develop for 8-10 days under conditions of 26℃-29℃ and 65%-75% relative humidity. This step provides a suitable growth environment for the parasitized wasp larvae, promoting their pupation and emergence.

[0013] Furthermore, after the tussah silkworm pupae have completed their development, a pre-cooling and storage step is included: well-parasitized tussah silkworm pupae are selected and placed in a single layer, first pre-cooled in a cold storage room at 10℃-12℃ for 36-48 hours, and then transferred to a cold storage room at 3℃-5℃ for later storage. This gradient cooling pre-cooling and storage method can effectively pause bee development, extend storage time, and maintain bee viability, facilitating subsequent release or transfer as needed.

[0014] Furthermore, the breeding container is a shallow container, and each breeding container contains 100 female or male tussah silkworm pupae. The shallow container facilitates ventilation and even contact of the bee colony, and the density of 100 pupae per box has been proven in practice to ensure parasitism efficiency while avoiding poor development caused by overcrowding.

[0015] Compared with the prior art, the present invention has the following significant advantages: 1. Significantly improved parasitism rate: By separating male and female bees and introducing them in a differentiated manner, bee species are more rationally distributed to hosts of different sexes, making full use of the carrying advantages of female and male pupae and improving the overall parasitism rate.

[0016] The rate of rotten pupae was significantly reduced: Simultaneous removal of diseased and damaged pupae and standardized disinfection procedures reduced pathogen infection, and the reasonable amount of bee colony released prevented excessive parasitism, thus reducing the rate of rotten pupae.

[0017] Bee colony resource conservation: Bee colonies are precisely released based on the actual carrying capacity of male and female pupae, avoiding the problem of waste or insufficient bee colonies in traditional mixed inoculation.

[0018] Improved breeding efficiency: The increase in parasitism rate and bee colony utilization rate directly led to an increase in the quantity and quality of bees produced.

[0019] Shortened emergence cycle: Optimized temperature and humidity control and development conditions enable more synchronized development of bee species, shortening the adult emergence period by 2-3 days, which is conducive to centralized release and large-scale application. Detailed Implementation

[0020] The present invention will be further described in detail below with reference to specific embodiments. Obviously, the described embodiments are merely some, not all, of the embodiments of the present invention. All other embodiments obtained by those skilled in the art based on the embodiments of the present invention without creative effort are within the scope of protection of the present invention.

[0021] The process flow of this invention mainly includes pretreatment, male-female separation and screening, cleaning and disinfection, differential inoculation, parasitic culture, development, and pre-cooling storage. The operating parameters and conditions for each step are described in detail below through specific embodiments.

[0022] Example 1: Standard Optimized Breeding Process This embodiment details the standard optimized process flow of the present invention, with all parameters taken as the median value of the preferred range to demonstrate the best implementation effect.

[0023] (1) Pretreatment: Remove the silkworm pupae (cocoons) with shells from the cold storage and spread them out in the operating room to dry for 2.5 hours. At this time, the cocoon shells will become dry and hard due to moisture evaporation. Manually peel the cocoons and place the naked pupae in a flat plastic basket for storage at room temperature for later use. Drying makes the cocoon shells dry and hard, reducing the difficulty of peeling the cocoons, reducing the risk of damage to the pupae, and providing a complete and healthy source of insects for subsequent steps.

[0024] (2) Sexing and screening: The tussah silkworm pupae were manually separated into males and females based on their external morphological characteristics (such as body size and abdominal rings). During the separation process, damaged and diseased pupae were picked out and placed in a special waste container. After identification, healthy female and male pupae were placed in shallow plastic baskets marked "female" and "male" respectively. Diseased and damaged individuals were removed simultaneously, effectively blocking the transmission route of pathogenic microorganisms and significantly reducing the phenomenon of rotten pupae caused by later group infection.

[0025] (3) Cleaning and disinfection: Place the sexed silkworm pupae into a special cleaning container and rinse them repeatedly with running water to remove cocoon silk, hemolymph residue, and dirt attached to their bodies. After removing them, let them air dry naturally, then spray or soak them in a 0.1% sodium hypochlorite solution for surface disinfection, followed by a second drying. This step further kills potential pathogens on the body surface, creating a clean environment for bee parasitism.

[0026] (4) Packing for inoculation: Place the sterilized and dried female and male tussah silkworm pupae into inoculation boxes, with 100 pupae per box. The shallow container design ensures that the pupae are distributed in a single layer, which is conducive to the bee colony making even contact with the host.

[0027] (5) Differentiated Inoculation: Move the inoculation boxes into the inoculation room. Inoculate in the order of female first, then male: Inoculate the first number of *Apis zhouensis* larvae into the female *tussah* pupa boxes, and the second number of larvae into the male *tussah* pupa boxes. The number of larvae per female *tussah* pupa is greater than the number of larvae per male *tussah* pupa. Specifically, using the average number of larvae per female *tussah* pupa during mixed inoculation as a reference, the number of larvae per female *tussah* pupa is higher than this average, and the number of larvae per male *tussah* pupa is lower than this average. In actual production, the specific number of larvae can be slightly adjusted according to the viability of the larvae and the quality of the host, but the relationship of the number of larvae per female *tussah* pupa being greater than that of male *tussah* pupa is always maintained.

[0028] (6) Parasitic Culture: After inoculation, seal and compact the inoculation box to prevent the bee colony from escaping. Move it into a light-free breeding room, adjust the temperature to 26.5℃ (midpoint of 25℃-28℃), and the relative humidity to 65% (midpoint of 60%-70%), and store it for 42 hours (midpoint of 36-48 hours). Maintain a slight airflow in the breeding room. This temperature and humidity combination is most suitable for the parasitic activities of *Aegilops zhouensis*, and the slight airflow ensures fresh indoor air and avoids mold growth caused by excessive humidity.

[0029] (7) Development: After parasitism is complete, open the breeding container and place the parasitized silkworm pupae in a single layer in a flat basket, then move them to the development room. Regulate the temperature in the development room to 27.5℃ (midpoint of 26℃-29℃) and the relative humidity to 70% (midpoint of 65%-75%), allowing development for 9 days (midpoint of 8-10 days). During this stage, the bee larvae feed and develop inside the pupae; the optimized temperature and humidity promote normal pupation and emergence.

[0030] (8) Pre-cooling and storage: After development, the healthy parasitized silkworm pupae are manually selected (excluding those that are not parasitized or infected) and placed in a single layer in a flat basket. They are first moved to a cold storage room at 10℃-12℃ for 42 hours (the midpoint of 36-48 hours) to allow the pupae to gradually adapt to the low temperature; then they are moved to a cold storage room at 3℃-5℃ for long-term storage. Gradual cooling avoids cold shock to the parasitized wasps caused by sudden temperature drops, effectively suspends the development process, prolongs the storage period, and maintains the high vitality of the bee species.

[0031] Example 2: High-Temperature Rapid Propagation Process Based on Example 1, this embodiment adjusts some parameters to adapt to high-temperature environments in summer or scenarios where it is necessary to accelerate the breeding cycle, demonstrating the flexible application of the present invention within the parameter range.

[0032] (1) Pretreatment: The drying time is shortened to 2 hours, so that the cocoon shell can be peeled, thus reducing indoor energy consumption.

[0033] (2) Separation and screening of male and female pupae: Same as in Example 1, strictly remove damaged and diseased pupae.

[0034] (3) Cleaning and disinfection: Same as in Example 1.

[0035] (4) Packaging: 100 heads per box, sorted by gender.

[0036] (5) Differentiated inoculation: Same as in Example 1, inoculate females first and then males, and the inoculation amount should be greater than that of males. The average inoculation amount is the same as in Example 1, but according to the characteristics of bee species with strong activity in the high temperature season, the deviation between the inoculation amount of females and males and the average value can be appropriately reduced to avoid energy consumption caused by excessive activity of bees.

[0037] (6) Parasitic culture: The temperature is set at 28℃ (upper limit of range), the relative humidity is 60% (lower limit of range), and the storage time is 36 hours (lower limit of range). The higher temperature stimulates the bee species to lay eggs and parasitize quickly, while the lower humidity and shorter time help prevent the growth of miscellaneous bacteria in the high temperature and high humidity environment, and at the same time adapt to the rapid metabolic needs of the bee species under high temperature.

[0038] (7) Development: The temperature was set at 29℃ (upper limit of range), the relative humidity at 65% (lower limit of range), and the development period was 8 days (lower limit of range). The higher temperature and slightly lower humidity accelerated the development of the bee colony, shortened the development cycle, and met the need for rapid bee emergence.

[0039] (8) Pre-cooling storage: Pre-cool at 12℃ for 36 hours, then move to 5℃ for storage. A higher initial pre-cooling temperature is suitable for a smooth transition from a higher development temperature, avoiding excessive temperature difference that could cause stress to the bee species.

[0040] Example 3: Low-temperature energy-saving breeding process This embodiment adjusts parameters to adapt to winter or energy-saving needs, demonstrating effective breeding under lower temperature conditions.

[0041] (1) Pretreatment: The drying time is extended to 3 hours to ensure that the cocoon shell is fully dry and easy to peel.

[0042] (2) Separation and screening of males and females: Same as in Example 1.

[0043] (3) Cleaning and disinfection: Same as in Example 1.

[0044] (4) Packaging: 100 heads per box.

[0045] (5) Differentiated inoculation: Same as in Example 1, the inoculation amount should be greater for females than for males. In the cold season, when bee activity is weak, the deviation between the female inoculation amount and the average value can be appropriately increased to ensure that female pupae receive a sufficient amount of bee colonies.

[0046] (6) Parasitic culture: The temperature was set at 25℃ (lower limit of range), the relative humidity at 70% (upper limit of range), and the storage time was 48 hours (upper limit of range). The lower temperature and higher humidity extended the survival and parasitism time of the bee species, ensuring the success rate of parasitism.

[0047] (7) Development: The temperature was set at 26℃ (lower limit of range), the relative humidity at 75% (upper limit of range), and the development time was 10 days (upper limit of range). The lower temperature and higher humidity slowed down the development speed, but were beneficial to the full development and large size of individual bees.

[0048] (8) Pre-cooling storage: Pre-cool at 10°C for 48 hours, then transfer to 3°C for storage. The longer pre-cooling time allows the pupae to cool slowly and fully at low temperatures, reducing the risk of chilling injury.

[0049] Comparative Example 1: Traditional Mixed Inoculation Method This comparative study used the traditional method for breeding Zhou's parasitic wasps, without separating males and females, and used mixed-sex tussah silkworm pupae in all steps.

[0050] (1) Pretreatment: Same as in Example 1, sun-dry for 2.5 hours, peel off the cocoons to obtain naked pupae.

[0051] (2) Screening: Simply remove obviously damaged pupae without systematically removing diseased ones.

[0052] (3) Cleaning and disinfection: Same as in Example 1.

[0053] (4) Packing: Place the mixed-sex tussah silkworm pupae into an inoculation box, 100 pupae per box.

[0054] (5) Inoculation: According to the conventional ratio for mixed inoculation, bee colonies are added to the box at one time. The average number of bee colonies added per tussah silkworm pupa is M. Since the sex is not distinguished, all pupae receive the same number of bee colonies.

[0055] (6) Parasitic culture: Same as in Example 1, 26.5℃, 65%RH, 42 hours.

[0056] (7) Development: Same as in Example 1, 27.5℃, 70%RH, 9 days.

[0057] (8) Storage: Same as in Example 1.

[0058] This ratio represents the current level of technology, but because it does not distinguish between sexes, bee species are randomly assigned during parasitism, resulting in insufficient parasitism of some female pupae and excessive parasitism of some male pupae, leading to low overall efficiency.

[0059] Comparative Example 2: Only males and females were separated, but they were planted in equal proportions. This comparative study, based on the separation of males and females, introduced the same proportion of bee species into female and male pupae to verify the necessity of differentiated bee introduction.

[0060] Steps (1) to (4): exactly the same as in Example 1, male and female separation, screening, disinfection and packaging by sex.

[0061] (5) Inoculation: Inoculation was carried out in the order of females first and then males, but the amount of bees put into the female boxes and male boxes was equal to the average number of bees put into the mixed inoculation, that is, the number of females and males put into the same box was equal. No differentiation was reflected.

[0062] Steps (6) to (8): Same as in Example 1.

[0063] This comparative test examined the effects of separating males and females, but ignored the differences in the carrying capacity of pupae of different sexes, thus failing to maximize reproductive benefits.

[0064] Effect verification and comparative analysis To visually demonstrate the technical advantages of this invention, the inventors conducted large-scale field comparative trials and compiled the breeding effect data of the three embodiments and two comparative examples, as shown in Table 1. The above data are based on parallel trials conducted by the inventors on the same batch of tussah silkworm pupae under identical environmental conditions, ensuring the comparability and reliability of the data.

[0065] Table 1 Comparison of the effects of different breeding methods Example 1 95.2 0.8 25 32.5 2.5 Example 2 93.8 0.9 24.2 31 3 Example 3 94.5 0.85 24.8 31.8 2 Comparative Example 1 65 5 0 0 0 Comparative Example 2 78.5 2.5 0 12.5 1 As can be seen from the data in Table 1: The parasitism rates of Examples 1 to 3 were all stable at over 93%, which was more than 30% higher than the 65% of Comparative Example 1 (traditional method) and also significantly improved compared to the 78.5% of Comparative Example 2, proving that the combination of male and female separation and differentiated seeding can greatly improve parasitism efficiency.

[0066] Regarding the rate of rotten pupae, the example controlled it at 0.8%-0.9%, which is much lower than 5% in Comparative Example 1 and 2.5% in Comparative Example 2, verifying the important role of removing diseased and damaged pupae, disinfection, and reasonable seeding in reducing the rate of rotten pupae.

[0067] The bee colony saving rate was only reflected in the example group, reaching 24%-25%, indicating that differentiated bee colony input avoided bee colony waste.

[0068] The improvement in efficiency was also evident only in the example group, with an increase of more than 31%, demonstrating the comprehensive economic advantages of the present invention.

[0069] Regarding the shortening of the emergence period, the example group showed a significant reduction of 2-3 days compared to the control group, which helps in the concentrated emergence and release of bee species.

[0070] In summary, this invention solves the technical problems of low parasitism rate, high pupa rot rate, and waste of bee colonies in traditional breeding methods by separating males and females, differentiating seeding, removing diseased and damaged bees, standardizing disinfection, and controlling temperature and humidity. It achieves efficient and high-quality breeding of *Aegilops zhouensis*, providing sufficient natural enemy resources for biological control.

Claims

1. A method for breeding *Pterocarya zhouensis* based on the separate inoculation of male and female *Tetraphyta tussah* pupae, characterized in that, Includes the following steps: The male and female tussah silkworm pupae were separated to obtain female and male tussah silkworm pupae; The female and male tussah silkworm pupae were placed in separate breeding containers. A first number of *Apis zhouensis* species are placed into a breeding container containing the female *tussah silkworm* pupae, and a second number of *Apis zhouensis* species are placed into a breeding container containing the male *tussah silkworm* pupae; wherein the number of species placed for a single female *tussah silkworm* pupae is greater than the number of species placed for a single male *tussah silkworm* pupae. After inoculation, the breeding containers were sealed and transferred to the breeding room for parasitic culture.

2. The breeding method according to claim 1, characterized in that, Before separating the male and female tussah silkworm pupae, the process includes pre-treatment of the tussah silkworm pupae with their shells on: placing the tussah silkworm pupae with their shells on indoors to dry for 2-3 hours, and then peeling off the cocoons after the cocoons have dried and hardened to obtain the tussah silkworm pupae.

3. The breeding method according to claim 1, characterized in that, During the process of separating male and female tussah silkworm pupae, damaged and diseased pupae are removed simultaneously, while healthy and intact tussah silkworm pupae are retained.

4. The breeding method according to claim 1, characterized in that, Before placing the female and male tussah silkworm pupae into separate breeding containers, a cleaning and disinfection step is included: the separated female and male tussah silkworm pupae are cleaned with water, dried, and then surface disinfected with disinfectant.

5. The breeding method according to claim 1, characterized in that, The step of releasing a first number of *Ariocarpus zhouensis* species into a breeding container containing the female *Tetracentron sinense* pupae, and releasing a second number of *Ariocarpus zhouensis* species into a breeding container containing the male *Tetracentron sinense* pupae, specifically includes: Following the inoculation order of females first and then males, the Zhou's biting wasp species were placed into the corresponding breeding containers. Taking the average number of wasp species per tussah silkworm pupa during mixed inoculation as a reference, the number of wasp species per female tussah silkworm pupa was higher than the average number, while the number of wasp species per male tussah silkworm pupa was lower than the average number.

6. The breeding method according to claim 1, characterized in that, The parasitic culture steps specifically include: After inoculation, the breeding container is sealed and compacted, and then moved into a breeding room with no light and slight air circulation. The temperature in the breeding room is controlled at 25℃-28℃ and the relative humidity at 60%-70%. The container is stored for 36-48 hours to allow the Zhou's parasitic wasp species to complete the parasitism of the tussah silkworm pupa.

7. The breeding method according to claim 1, characterized in that, After the parasitic culture is completed, the process also includes a development step: the parasitized silkworm pupae are placed in a single layer and moved to a development room, where they develop for 8 to 10 days under conditions of 26℃-29℃ and 65%-75% relative humidity.

8. The breeding method according to claim 7, characterized in that, After the tussah silkworm pupae have completed their development, a pre-cooling and storage step is also included: select well-parasitized tussah silkworm pupae and place them in a single layer. First, move them to a cold storage room at 10℃-12℃ for pre-cooling for 36-48 hours, and then move them to a cold storage room at 3℃-5℃ for storage and future use.

9. The breeding method according to claim 1, characterized in that, The breeding container is a shallow container, and each breeding container contains 100 female or male tussah silkworm pupae.